Enhanced luminescent or luminometric assay

• Main Authors: KRICKA; LARRY J, THORPE; GARY H. G. H, WHITEHEAD; THOMAS P, O'TOOLE; ANGELA M
• Format: Patent
• Language: English
• Published: 08.03.1988
• Edition: 4
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PHYSICS; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPIRITS; TESTING; VINEGAR; WINE

It has been a problem that the chemiluminescent emission produced by the peroxidase-catalyzed oxidation of a 2,3-dihydro-1,4-phthalazinedione is often weak or gives a poor signal-background ratio and therefore does not give the high degree of sensitivity required for assay purposes, particularly for assay of peroxidase used to label a reagent for example in ELISA. The present invention solves the problem by the finding the certain narrowly defined aromatic amines are enhancers of the reaction and accordingly provides for their use in an assay and kit. The amines are of general formula (I) (1) wherein the R symbols (that is, R, R1, R2, R3, R4, R5 and R6) have any of the meanings (a) to (j) given below, all other R symbols in each meaning are hydrogen atoms and the fused rings are to be read in the same configurational sense as formula I: (a) R=R1=CH3; R4= (b) R4= cyclohexyl, or alkyl or alkoxy having 1 to 4 carbon atoms (c) R2, R3 and R4 together represent the polycyclic fused ring system: (d) R3 and R4 together represent the fused ring or ring system (e) R3 and R4 together represent the fused ring system and R5 and R6 together represent the fused ring (f) R2=NH2 and R4= (g) R2=R6=CH3 and R4= (h) R3=C2H5 (i) R2=R4=CH3 and (j) R3=R4=CH3.